The invention belongs to the technical field of thermo-printing material and discloses an anti-counterfeiting bar code printing thermal transfer ribbon and a preparation method thereof. The anti-counterfeiting bar code printing thermal transfer ribbon is composed of a base ribbon, and a back coating and a thermal transferring coating which are respectively coated on two sides of the base ribbon; the thermal transferring layer is divided into two parts, the surface at one side of the base ribbon is sequentially provided with a separating layer and a color layer from inside to outside, and the color layer with fluorescent material is made by the following raw materials by weight percentage: 35-68 percent of binders, 20-35 percent of waxes, 5-15 percent of pigment, 5-15 percent of fluorescentmaterial and 2-3 percent of assistants. The back coating is coated at one side of the base ribbon, and then a separating layer and the color layer are coated on the other side of the base ribbon, soas to form the anti-counterfeiting bar code printing thermal transfer ribbon. The anti-counterfeiting bar code printing thermal transfer ribbon integrates anti-counterfeiting ink and thermal transferring bar code technology and overcomes the problem that large-batch anti-counterfeiting ink printing has high cost, thereby meeting the requirements of classification of goods and enhancing the security and safety of goods.